One problem that exists in the art today is that there remains a lack of methods and systems that both improve detection of abnormalities in biological samples and deliver analytical results to a practitioner.
In the related art, a number of diseases may be diagnosed using classical cytopathology and histopathology methods involving examination of nuclear and cellular morphology and staining patterns. Typically, such diagnosis occurs via examining up to 10,000 cells in a biological sample and finding about 10 to 50 cells or a small section of tissue that may be abnormal. This finding is based on subjective interpretation of visual microscopic inspection of the cells in the sample.
An example of classical cytology dates back to the middle of the last century, when Papanicolaou introduced a method to monitor the onset of cervical disease by a test, commonly known as the “Pap” test. For this test, cells are exfoliated using a spatula or brush, and deposited on a microscope slide for examination. In the original implementation of the test, the exfoliation brush was smeared onto a microscope slide, hence the name “Pap smear.” Subsequently, the cells were stained with hematoxylinleosin (H&E) or a “Pap stain” (which consists of H&E and several other counterstains), and were inspected visually by a cytologist or cyto-technician, using a low power microscope.
The microscopic view of such samples often shows clumping of cells and contamination by cellular debris and blood-based cells (erythrocytes and leukocytes/lymphocytes). Accordingly, the original “Pap-test” had very high rates of false-positive and false-negative diagnoses. Modern, liquid-based methods (such as cyto-centrifugation, the ThinPrep® or the Surepath® methods) have provided improved cellular samples by eliminating cell clumping and removing confounding cell types.
However, although methods for the preparation of samples of exfoliated cells on microscope slides have improved substantially, the diagnostic step of the related art still typically relies on visual inspection and comparison of the results with a data base in the cytologist's memory. Thus, the diagnosis is still inherently subjective and associated with low inter- and intra-observer reproducibility. To alleviate this aspect, other related art automated visual light image analysis systems have been introduced to aid cytologists in the visual inspection of cells. However, since the distinction between atypia and low grades of dysplasia is extremely difficult, such related art automatic, image-based methods have not substantially reduced the actual burden of responsibility on the cytologist.
In classical histopathology, tissue sections, rather than exfoliated individual cells, are inspected by a pathologist using a microscope after suitable staining of the tissue. To detect abnormalities, the pathologist focuses on gross tissue architecture, cell morphology, nuclear morphology, nucleus-to-cytoplasm ratio, chromatin distribution, presence of mitotic figures, and others. Since these criteria are morphology-based, their interpretation always will be somewhat subjective. Immuno-histochemical and other more recent methods are often used to augment the pathologist's subjective assessment of a tissue diagnosis.
Spectral methods have also been applied in the related art to the histopathological diagnosis of tissue sections available from biopsy. The data acquisition for this approach, referred to as “Spectral Histopathology (SHP),” can be carried out using the same spectral methodology used for spectral cytopathology (“SCP”).
In some methods of the related art, a broadband infrared (IR) or other light output is transmitted to a sample (e.g., a tissue sample), using instrumentation, such as an interferometer, to create an interference pattern. Reflected and/or transmitted light is then detected, typically as an interference pattern. A Fast Fourier Transform (FFT) may then be performed on the detected pattern to obtain spectral information relating to each sample pixel. The resulting information is referred to as a pixel spectrum.
One limitation of the FFT based related art process is that the amount of radiative energy available per unit time in each band pass may be very low, due to use of a broadband infrared spectrum emission. As a result, the data available for processing with this approach is generally inherently noise limited. Further, in order to discriminate the received data from background noise, for example, with such low energy levels available, high sensitivity instruments must be used, such as high sensitivity liquid nitrogen cooled detectors (the cooling alleviates the effects of background IR interference). Among other drawbacks, such related art systems may incur great costs, footprint, and energy usage.
Thus, there remains a need in the art for devices, systems, and methods for analyzing IR and/or other similar transmissions for identifying cellular, tissue, biochemical, molecular and morphologic subtypes in an image of a biological specimen for diagnosis, prognosis, therapies and/or prediction of diseases and/or conditions.